This study was conducted to investigate the anti-diabetic activity of Kocat-D1, which is widely used in traditional medicine to treat diabetes in Shandong, China. Sprague Dawley rats (8 weeks of age) were separated into 4 groups: a normal control, streptozotocin (STZ)-induced diabetic rat group (DM control), Kocat-D1-1 (diabetic rat treated with 0.25 g/kg/day hot water extract), and Kocat-D1-2 (diabetic rat treated with 1 g/kg/day hot water extract). After eight weeks of treatment, the fasting blood glucose levels of the Kocat-D1-1 ($334.3{\pm}32.9\;mg/dL$) and Kocat-D1-2 group ($259.5{\pm}35.0\;mg/dL$) were significantly lower when compared to the DM control group ($451{\pm}42.6\;mg/dL$). Furthermore, the levels of glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT), albumin and high-density lipoprotein (HDL) cholesterol in the serum of the Kocat-D1-2 group were significantly normalized when compared to the DM control group. However, significant differences were not observed between the Kocat-D1-1 group and the DM control group. Histochemical staining of the liver of the Kocat-D1-2 group revealed no fat accumulation. The insulin level was significantly upregulated in the Kocat-D1-2 group ($0.13{\pm}0.02\;ng/mL$) when compared to the DM control group ($0.05{\pm}0.04\;ng/mL$). The relative volume of $\beta$-cells in the pancreas of the Kocat-D1-2 group ($49.4{\pm}4.2%$) also increased significantly when compared to the DM control group ($12.9{\pm}7.9%$). These results suggest that Kocat-D1 exerts an anti-hyperglycemic effect through the enhancement of insulin secretion.
Background: Luteolin, a flavone found in various Chinese herbal medicines is known to possess anti-inflammatory properties through its ability to inhibit various proinflammatory signaling pathways including NF-${\kappa}B$ and p38 MAPK. In this study, we investigated the potential therapeutic effect of luteolin on dextran sodium sulfate (DSS)-induced colitis. Materials and Methods: We used a transgenic mouse model expressing the enhanced green fluorescent protein (EGFP) under the transcriptional control of NF-${\kappa}B$$cis$-elements. C57BL/6 NF-${\kappa}B^{EGFP}$ mice received 2.5% DSS in their drinking water for six days in combination with daily luteolin administration (1mg/kg body weight, 0.1ml vol, intragastric) or vehicle. NF-${\kappa}B$ activity was assessed macroscopically with a Charge-Coupled Device (CCD) camera and microscopically by confocal analysis. Results: A significant increase in the Disease Activity Index (DAI), histological score (p<0.05), IL-12 p40 secretion in colonic stripe culture (p<0.05) and EGFP expression was observed in luteolin and/or DSS-treated mice compared to water-treated mice. Interestingly, a trend toward a worse colitis (DAI, IL-12p40) was observed in luteolin-treated mice compared to non-treated DSS-exposed mice. In addition, EGFP expression (NF-${\kappa}B$ activity) strongly increased in the luteolin-treated mice compared to control mice. Confocal microscopy showed that EGFP positive cells were primarily lamina propria immune cells. Conclusions: These results suggest that luteolin is not a therapeutic alternative for intestinal inflammatory disorders derived for primary defects in barrier function. Thus, therapeutic intervention targeting these signaling pathways should be viewed with caution.
Shin, Ji Eun;Lee, Kyungmin;Kim, Ji-Hee;Madhi, Iskander;Kim, YoungHee
Journal of Life Science
/
v.29
no.4
/
pp.402-409
/
2019
Korean red ginseng made from steaming and drying fresh ginseng has long been used as a traditional herbal medicine due to its effects on the immune, endocrine, and central nerve systems and its anti-inflammatory activity. In this study, we investigated the molecular mechanism responsible for the anti-inflammatory effects of a formulated Korean red ginseng extract (RGE) in response to lipoteichoic acid (LTA), a cell wall component of gram-positive bacteria. RGE inhibited LTA-induced nitric oxide (NO) secretion and inducible nitric oxide synthase (iNOS) expression in BV-2 microglial cells, without affecting cell viability. RGE also inhibited nuclear translocation of nuclear factor kappa B ($NF-{\kappa}B$) p65 and degradation of $I{\kappa}B-{\alpha}$. In addition, RGE increased the expression of heme oxygenase-1 (HO-1) in a dose-dependent manner, and the inhibitory effect of RGE on iNOS expression was abrogated by small interfering RNA-mediated knockdown of HO-1. Moreover, RGE induced nuclear translocation of nuclear factor E2-related factor 2 (Nrf2), a transcription factor that regulates HO-1 expression. Furthermore, the phosphoinositide-3-kinase (PI-3K) inhibitor and mitogen-activated protein kinase (MAPK) inhibitors suppressed RGE-mediated expression of HO-1, and RGE enhanced the phosphorylation of Akt, extracellular signal-regulated kinases (ERKs), p38, and c-JUN N-terminal kinases (JNKs). These results suggested that RGE suppressed the production of NO, a proinflammatory mediator, by inducing HO-1 expression via PI-3K/Akt- and MAPK-dependent signaling in LTA-stimulated microglia. The findings indicate that RGE could be used for the treatment of neuroinflammation induced by grampositive bacteria and that it may have therapeutic potential for various neuroinflammation-associated disorders.
With strong biotin binding affinity ($K_D=10^{-14}M$), the tetrameric feature of streptavidin could be used to increase the antigen binding activity of a camel heavy chain (VHH) antibody through their fusion, here stained with biotinylated horseradish peroxidase and subsequent immunoassays ELISA and Western blot analysis. For this application, we cloned the streptavidin gene amplified from the Streptomyces avidinii chromosome by PCR, and this was fused to the gene of the 8B9 VHH antibody which is specific to green fluorescent protein (GFP) antigens. To express a soluble fusion protein in Escherichia coli, we used the pUC119 plasmid-based expression system which uses the lacZ promoter for induction by IPTG, the pelB leader sequence at the N-terminus for secretion into the periplasmic space, and six polyhistidine tags at the C-terminus for purification of the expressed proteins using an $Ni^+$-NTA-agarose column. Although streptavidin is toxic to E. coli because of its strong biotin binding property, this soluble fusion protein was expressed successfully. In SDS-PAGE, the size of the purified fusion protein was 122.4 kDa in its native condition and 30.6 kDa once denatured by boiling, suggesting the tetramerization of the monomeric subunit by non-covalent association through the streptavidin moiety fusing to the 8B9 VHH antibody. In addition, this fusion protein showed biotin binding activity similar to streptavidin as well as GFP antigen binding activity through both ELISA and Western blot analysis. In conclusion, the protein resulting from the fusion of an 8B9 VHH antibody with streptavidin was successfully expressed and purified as a soluble tetramer in E. coli; it showed both biotin and GFP antigen binding activity suggesting the possible production of a tetrameric and bifunctional VHH antibody.
Purpose : The purpose of this study was to investigate the temperature changes of the ocular surface before and after cataract surgery using thermography of a thermal imaging camera. Methods : The study included 75 patients (75 eyes) aged from 50 to 79 years who underwent cataract surgery. In the past, those who underwent corneal-related surgery, wearing contact lens, disorder of tear secretion and taking medication for systemic disease were excluded from this study. The temperature changes of the eyeball surface were measured using a thermal imager (Cox CX series, Answer, Korea) following Tear Break Up Time (TBUT) test, Mcmonnies questionnaire and Schirmer's Test in real time, Results : While the temperature of preoperative ocular surface was $35.20{\pm}0.54^{\circ}C$ and that of postoperative temperature was $35.30{\pm}0.53^{\circ}C$, the difference was not significant. The temperature changes in the ocular surface were statistically significant at $-0.12{\pm}0.08{\Delta}$ ($^{\circ}C/sec$) before the surgery and $-0.18{\pm}0.07{\Delta}$ ($^{\circ}C/sec$) after the surgery. In comparison of the age groups, it was shown that the changes in the surface temperature before the surgery were from $-0.19{\pm}0.05{\Delta}$ ($^{\circ}C/sec$) to $-0.14{\pm}0.09{\Delta}$ ($^{\circ}C/sec$) in the 50s group, and from $-0.12{\pm}0.08{\Delta}$ ($^{\circ}C/sec$) to $-0.15{\pm}0.07{\Delta}$ ($^{\circ}C/sec$) in 60s group, and $-0.18{\pm}0.07{\Delta}$ ($^{\circ}C$) to $-0.12{\pm}0.08{\Delta}/sec$) in the 70s group, showing significant changes in the ocular surface temperature at all ages. Conclusion : Following the cataract surgery, all the indicators of dry eye syndrome were decreased, and eye surface temperature changes were significant. The thermography technique of the ocular surface would be expected to be useful for the evaluation of various dry eye syndromes because it is easy to evaluate dry eye syndrome noninvasively and can be quantified.
Background: The leaves and roots of Panax ginseng are rich in ginsenosides. However, the chemical compositions of the leaves and roots of P. ginseng differ, resulting in different medicinal functions. In recent years, the aerial parts of members of the Panax genus have received great attention from natural product chemists as producers of bioactive ginsenosides. The aim of this study was the isolation and structural elucidation of novel, minor ginsenosides in the leaves of P. ginseng and evaluation of their antiinflammatory activity in vitro. Methods: Various chromatographic techniques were applied to obtain pure individual compounds, and their structures were determined by nuclear magnetic resonance and high-resolution mass spectrometry, as well as chemical methods. The antiinflammatory effect of the new compounds was evaluated on lipopolysaccharide-stimulated RAW 264.7 cells. Results and conclusions: Two novel, minor triterpenoid saponins, ginsenoside $LS_1$ (1) and 5,6-didehydroginsenoside $Rg_3$ (2), were isolated from the leaves of P. ginseng. The isolated compounds 1 and 2 were assayed for their inhibitory effect on nitric oxide production in LPS-stimulated RAW 264.7 cells, and Compound 2 showed a significant inhibitory effect with $IC_{50}$ of $37.38{\mu}M$ compared with that of NG-monomethyl-L-arginine ($IC_{50}=90.76{\mu}M$). Moreover, Compound 2 significantly decreased secretion of cytokines such as prostaglandin $E_2$ and tumor necrosis factor-${\alpha}$. In addition, Compound 2 significantly suppressed protein expression of inducible nitric oxide synthase and cyclooxygenase-2. These results suggested that Compound 2 could be used as a valuable candidate for medicinal use or functional food, and the mechanism is warranted for further exploration.
Our previous studies have reported that antimicrobial peptides (AMPs) derived from the larvae of white-spotted flower chafer (Protaetia brevitarsis seulensis) exert anti-inflammatory and neuroprotective activities. This study explored the anti-inflammatory effects of protaetiamycine 9 (CVLKKAYFLTNLKLRG-NH2), a novel AMP, derived from P. b. seulensis against lipopolysaccharide (LPS)-mediated inflammatory response in RAW264.7 macrophage cells. Protaetiamycine 9 (25, 50, 75, and 100 ㎍/ml) did not cause cytotoxic effects against RAW264.7 cells. The RAW264.7 cells were pre-treated with various concentrations of protaetiamycine 9 (25-100 ㎍/ml) for 1 hr and then exposed to LPS (100 ng/ml) for 24 hr. Protaetiamycine 9 treatments decreased the LPS-induced secretion of inflammatory mediators, such as nitric oxide (NO), in a dose-dependent manner. Protaetiamycine 9 (25-100 ㎍/ml) effectively downregulated the LPS-induced increase in mRNA and the protein expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2), which are involved in the production of inflammatory mediators. Protaetiamycine 9 also suppressed the production and gene expression of pro-inflammatory cytokines, including interleukin (IL)-6 and IL-1β, compared to the presence of LPS alone. Furthermore, protaetiamycine 9 inhibited the degradation of inhibitory kappa B alpha (IκB-α) and the phosphorylation of mitogen-activated protein kinases (MAPKs), such as extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38. In conclusion, these results suggest that protaetiamycine 9 exhibits LPS-mediated inflammatory responses by blocking IκB-α degradation and MAPK phosphorylation.
There has been much debate on the morphometric divergence between the recently identified Apis cerana koreana and Apis cerana honey bees. The aim of this study was to obtain phenotypic information that can be used to compare A. c. koreana data with other A. cerana subspecies data from open resources and determine breeding results on the basis of morphometric traits. To differentiate A. c. koreana, we investigated 22 classic morphological characteristics; royal jelly secretion; and the weight of workers, queens, and drones of A. c. koreana bred in Korea. To define the selection results, we used the geometric morphometric method. The artificially selected A. c. koreana secreted significantly more royal jelly (1.18 times) than the naturally selected A. c. koreana, which positively influenced the health of the colonies. These honey bees were identified more clearly with the geometric morphometric method than with the classic morphometric method, which is traditionally used to determine the subspecies. Large trends were noted for A. c. koreana on the basis of our results and literature from the 1980s regarding A. cerana sizes in Korea (tarsal index, length of forewing, and cubital index were measured). The cluster analysis revealed the proximity of A. c. koreana, A. cerana in China, and A. c. indica on the basis of eight classic characters, which, perhaps, relay the origin of the honey bees. The results of this study defined the morphometric responses of A. c. koreana honey bees to geographic isolation, climate change, and selection, which are important to identify, protect, and preserve honey bee stock in Korea.
This study presented a measure for turning by-products, released from land farming sites, into resources. The measure involved adding food by-products such as rice bran and nonfat soybean to the sludge, released from the eel farming sites, inoculating the lactic acid bacteria, Aspergillus oryzae, and Bacillus subtilis by step, fermenting them, and measuring the changed ingredients of the fermented fodder. The water content of the fermented fodder by the step of preparation was the first-step fermented product (14.6%) using the lactic acid bacteria, and the second and third-stage fermented product (33.0% and 34.0% respectively) using Aspergillus oryzae and Bacillus subtilis. The pH level was found to be 5.38 in the first-step fermented product due to the secretion of lactic acid caused by the lactic acid bacteria, and the pH level of the second and third-stage fermented products was 5.66 and 7.26, respectively, showing that the pH level increased. The phytic acid content was 0.126g/100g in the first-step fermented product, 0.004g/100g in the second-stage fermented product, and 0.093g/100g in the third-stage fermented product. The measurement of nitrogen content revealed that the amino nitrogen content was high with 1226.37mg% in the second-stage fermented product, and a little lower with 710.18mg% in the third-stage fermented product. The ammonium nitrogen content increased from 0.988mg/kg in the first-stage fermented product to 1.502mg/kg in the third-stage fermented product. Total nitrogen content increased to 2.78% in the first-stage fermented product, 4.08% in the second-stage fermented product, and 4.85% in the third-stage fermented product. As fermentation continued with the three microbes, the phytic acid decreased, and the protein decomposition rate increased. Also, due to the 3 step fermentation, the low-molecule nitrogen ingredient content increased, suggesting that the fodder was developed to offer high digestion and absorption.
Young Ik Lee;Ahtesham Hussain;Md Aziz Abdur Rahman;Ho Yong Sohn;Hye Jung Yoon;Jin Sook Cho
Journal of Life Science
/
v.33
no.11
/
pp.923-935
/
2023
Rubus crataegifolius (RC), Ulmus macrocarpa (UM), and Gardenia jasminoides (GJ) are well-known folk medicines in Asia used to treat various gastrointestinal disturbances. The present study evaluated the gastroprotective effect of LS-RUG-com, a mixture of commercially prepared powders of RC, UM, and GJ with a ratio of 3:1:2(w/w/w) against HCl/ethanol-induced gastritis, indomethacin-induced ulcers, and esophageal reflux-induced esophageal mucosal damage and Helicobacter pylori infections. In addition, TNF-α and IL-1β expressions were also determined and measured in esophageal tissue. As to HCl/ethanol-induced gastritis, the LS-RUG-com treatment at a dose of 150 mg/kg showed a remarkable anti-gastritis effect. Regarding indomethacin-induced gastric ulcers, the LS-RUG-com treatment had a significant anti-gastric ulcer effect. Furthermore, in the gastroesophageal reflux disease (GERD) model experiment, the LS-RUG-com treatment resulted in the histological recovery of stomach damage and mucosal injuries. Furthermore, the LS-RUG-com treatment led to an increase in gastric content pH, an increase in mucus protection, and a decrease in gastric pepsin output with a significant decrease in TNF-α and IL-1β. As to the Helicobacter pylori infected animal model, LS-RUG-com had a notable inhibitory effect on Helicobacter growth. The use of RC, UM, or GJ in isolation or the LS-RUG-com treatment as whole had good effects in terms of anti-oxidation, anti-neutralization, gastric acid secretion inhibition, and anti-lipid peroxidation, which supported the use of natural products as systemic gastric protective agents. Our results suggest that the LS-RUG-com might be a significant systemic gastroprotective agent that could be utilized for the treatment and/or protection from gastric disturbances and related damage.
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